The association between cytomegalovirus (CMV)-specific reactivity of T cell subsets and development of CMV retinitis (CMV-R) was prospectively studied in 50 CMV-seropositive AIDS patients. The frequency of CMV-specific CD69 expression on CD8 T cells was similar in patients with and patients without CMV-R (median, 1.0% vs. 1.2%; P=.14). However, the frequency of CMV-specific CD69 expression on CD4 T cells was significantly lower in patients with CMV-R than in those without CMV-R (median, 0.4% vs. 2.25%; P<.001). CMV-specific CD4 T cell reactivity in patients who developed CMV-R shortly after starting highly active antiretroviral therapy (HAART) remained low, although the CD4 cell counts increased markedly. Therefore, development of CMV-R is associated with a poor CMV-specific reactivity of CD4 T cells but not with poor reactivity of CD8 T cells. Development of CMV-R after initiation of HAART is associated with a poor reconstitution of CMV-specific immune response, rather than with immune rebound.